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Thompson, N. F.

Publications and source records attributed to Thompson, N. F..

2 recordsLinked to original sources

Pedigree-based genome-wide imputation using a low-density amplicon panel for the highly polymorphic Pacific oyster Crassostrea (Magallana) gigas

High-density genomic data are instrumental for selective breeding, but the costs associated with these approaches can hinder progress, as is the case for most aquaculture species. A strategy to reduce genotyping costs is to genotype a few select individuals at high-density (e.g., parents, grandparents), and many others at low density (e.g., offspring), then impute genotypes. This has been demonstrated in silico for Pacific oyster Crassostrea (Magallana) gigas but was particularly challenging relative to other species and has never been empirically tested. Here, four families of Pacific oysters, bred via marker-assisted selection for variation at a locus for field survival in an ostreid herpesvirus 1 (OsHV-1)-positive estuary, were exposed to OsHV-1 then genotyped using a low-density amplicon panel (n = 240 individuals). Parents were genotyped with the amplicon panel and by whole-genome resequencing. Offspring genotypes were imputed, and accuracy was determined by comparing against held-out whole-genome data for offspring. Imputation resulted in reduced minor allele frequencies and enriched homozygosity relative to empirical data. An in silico three-generation analysis was used to investigate the effect of deepening the pedigree, resulting in superior concordance in genotypes (GC = 84.5%) and allelic dosage (r = 0.73) compared to two-generation imputation (GC = 75.3%; r = 0.63). Genome-wide associations to OsHV-1 survivorship with imputed data identified significantly associated regions on the expected chromosome 8, but not at the expected position based on previous work, pointing to a potentially more complex genetic architecture for the trait. Our results empirically demonstrate the utility of amplicon panel-based genome-wide imputation in shellfish, and thus enable low-cost selective breeding techniques.

genomics↗

An amplicon panel for high-throughput and low-cost genotyping of Pacific oyster

Maintaining genetic diversity in cultured shellfish can be challenging due to high variance in individual reproductive success, founder effects, and rapid genetic drift, but is important to retain adaptive potential and avoid inbreeding depression. To support broodstock management and selective breeding in cultured Pacific oysters (Crassostrea (Magallana) gigas), we developed an amplicon panel targeting 592 genomic regions and SNP variants with an average of 50 amplicons per chromosome. Target SNPs were selected based on elevated observed heterozygosity or differentiation in Pacific oyster populations in British Columbia (BC), Canada. The use of the panel for parentage applications was evaluated by genotyping three generations of oysters from a breeding program in BC (n = 181) and a set of families that were selected for Ostreid herpesvirus-1 (OSHV-1) resistance from the Molluscan Broodstock Program, a Pacific oyster breeding program in Oregon, USA (n = 136). Population characterization was evaluated using collections of wild, naturalized, farmed, or hatchery oysters sampled throughout the Northern Hemisphere (n = 190). Technical replicate samples showed high genotype concordance (97.5%; n = 68 replicates). Initial parentage analysis found instances of suspected pedigree and sample handling errors, demonstrating the panels value for quality control in breeding programs. Suspected null alleles were identified in parentage datasets and were found to reduce assignment success. Null alleles were largely population dependent, suggesting population-specific variation impacts target amplification. By taking an iterative approach, null alleles were identified using existing data without the need for pedigree information, and once null alleles were removed, assignments increased to 93.0% and 86.0% of possible assignments in the two breeding program datasets. A pipeline for analyzing the amplicon sequence data from sequencer output, amplitools, is also provided.

genetics↗